title: Rectangle FRAP for measuring diffusion with a laser scanning microscope.
abstract: Fluorescence recovery after photobleaching (FRAP) is one of the most useful microscopy techniques for studying the mobility of molecules in terms of a diffusion coefficient. Here, we describe a FRAP method that allows such measurements, relying on the photobleaching of a rectangular region of any size and aspect ratio. We start with a brief overview of the rectangle FRAP theory, and next we provide guidelines for performing FRAP measurements, including a discussion of the experimental setup and the data analysis. Finally, we discuss how to verify correct use of the rectangle FRAP method using test solutions.
authors.affiliation: Laboratory of General Chemistry and Physical Pharmacy, Ghent University and Centre of Nano- and Biophotonics, Ghent, Belgium.
authors.forename: Ranhua
authors.initials: R
authors.lastname: Xiong
authors.forename: Hendrik
authors.initials: H
authors.lastname: Deschout
authors.forename: Jo
authors.initials: J
authors.lastname: Demeester
authors.forename: Stefaan C
authors.initials: SC
authors.lastname: De Smedt
authors.forename: Kevin
authors.initials: K
authors.lastname: Braeckmans
mesh_terms: D004058:Diffusion; D036681:Fluorescence Recovery After Photobleaching; D049452:Green Fluorescent Proteins / Q000737:chemistry; D018613:Microscopy, Confocal / Q000379:methods*; D012996:Solutions / Q000737:chemistry
chemical_list: D012996:Solutions; D049452:Green Fluorescent Proteins